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1500 PH Seminar Meirose
1500 PH Seminar Meirose
pH = - log aH
• In solutions that contain other ions, activity and concentration are not
the same. The activity is an effective concentration of hydrogen ions,
rather than the true concentration; it accounts for the fact that other
ions surrounding the hydrogen ions will shield them and affect their
ability to participate in chemical reactions.
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What is pH?
Representative pH values
Substance pH
• The pH Scale Hydrochloric Acid, 10M -1.0
Lead-acid battery 0.5
Gastric acid 1.5 – 2.0
• Each pH unit is a factor 10 in Lemon juice 2.4
[H+] Cola 2.5
Vinegar 2.9
• pH of Cola is about 2.5. This Orange or apple juice 3.5
is 10x more acidic than Beer 4.5
Orange Juice (pH of 3.5) Acid Rain <5.0
Coffee 5.0
Tea or healthy skin 5.5
• Cola is 100x more acidic Milk 6.5
than Beer! (pH of 4.5) Pure Water 7.0
Healthy human saliva 6.5 – 7.4
Blood 7.34 – 7.45
Seawater 7.7 – 8.3
Hand soap 9.0 – 10.0
Household ammonia 11.5
Bleach 12.5
Household lye 13.5
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pH Measurement System
• When two solutions containing different concentrations of H+ ions are
separated by a glass membrane, a voltage potential is developed
across the membrane. (Sensing electrode)
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pH Measurement System
• The pH Meter
• Acts as a volt meter
• Translates electrode potential (mV) to
pH scale
• Meter functions
• Stores calibration curve
• Adjusts for temperature changes
• Adjusts electrode slope
• Signals when reading is stable
• Features
• mV and relative mV scales
• Recognizes US Standard Buffers
• Number of calibration points
• Display information
• RS232 or recorder outputs
• Datalogging
• GLP/GMP compliant
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Measuring pH
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pH Measurement System
The pH Electrode
• Combination
• Sensing Half-Cell
• Reference Half-Cell
Common References
• Calomel (going, going…..)
• Ag/AgCl
• ROSS™
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pH Measurement System – Reference Electrode
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Common Questions – Electrode Types
• What is a triode?
• A triode is a combination electrode
(sensing and reference together) plus an
ATC (automatic temperature
compensation thermistor) all built into one
electrode body. (Same size as a
reference, sensing or combination
electrode.)
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pH Measurement System – Reference Types
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pH Measurement System – Reference Types
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pH Measurement System – Reference Types
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pH Measurement System – Reference Types
Disadvantages • Cost
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pH Measurement System - Junctions
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pH Measurement System - Junctions
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pH Measurement System - Junctions
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Common Questions – Electrode Types
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pH Measurement System – Electrode Types
Refillable or Low Maintenance Gel?
Low Maintenance Gel Electrodes
• Easy to use
• Rugged epoxy body
• 0.05-0.1 pH precision
• Slower response rate
• 6 month average life
• Gel memory effects at junction
Refillable Electrodes
• Fill/drain electrode
• Wide applicability
• Glass or epoxy body
• 0.02 pH precision
• Faster response rate
• 1 year minimum life
• Replaceable fill solution
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pH Measurement System – Electrode Types
Polymer or Low Maintenance Gel?
Low Maintenance Gel Electrodes
• Easy to use
• Rugged epoxy body
• 0.05-0.1 pH precision
• Slower response rate
• 6 month average life
• Gel memory effects at junction
Polymer Electrodes
• Low maintenance
• Easy to use
• Glass or epoxy body
• 0.02 pH precision
• Faster response rate
• 1 year minimum life
• Double junction design
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pH Measurement System - Electrode Selection
• Select proper reference for application
• ROSS™, Single or Double Junction Ag/AgCl
• Remember that Calomel contains Mercury!
• Select proper junction for application
• Wick, Ceramic, Open, Sure-Flow, etc.
E = E0 - RT/nF log aH
E = measured potential
E0 = reference potential
R = Universal Gas Constant
T= Temperature (at 25 oC)
n = Number of electrons
F = Faraday Constant
aH = Hydrogen Ion activity
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Walther Nernst
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Albert Einstein
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pH Calibration
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pH Calibration
• When you are calibrating, you are determining the electrodes slope as it
relates to the theoretical slope defined by the Nernst Equation
• Newer meters automatically calculate slope
• Check slope manually by reading mV in buffers and comparing to
Nernstian response (59.2 mV/pH unit)
• Example:
• pH 7 = -10 mV
• pH 4 = +150 mV
• Slope = 160 mV/177.6 mV = 90.1%
• Where did this 177.6mV come from?
− A change of 3 pH units (7-4)
− 59.2 mV per pH unit x 3 equals 177.6 mV
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Common Questions: Calibration
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Common Questions: Calibration
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pH Calibration - Guidelines
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Effects of Temperature
• Electrode
• Calibration
• Buffers
• Samples
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Effects of Temperature – Electrode Effects
• Temperature Hysteresis
• AgCl or Hg2Cl2 references
drift with temperature
changes
• 0.05 pH unit error with 4 oC
difference
• ROSS™ electrodes stabilize
within seconds
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Effects of Temperature – Calibration Effects
• Calibration Effects
• Theoretical slope of electrode is
59.16mv at 25 oC
• Temperature changes the
calibration slope
• Temperature compensation adjusts
the calibration slope for
temperature effects
• The point at which temperature has
no effect on mV is referred to as
the isopotential point
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Effects of Temperature – Buffer Effects
• Buffer Effects
• Buffers have different pH values at different temperatures
• Use the value of the buffer at the calibration temperature
• New meters have NIST calibration tables pre-programmed
• NIST Certified Values only at 25°C
25 C 0C 5C 10 C 20 C 30C 40 C 50 C 60 C 70 C 80 C 90 C
1.68 1.67 1.67 1.67 1.67 1.68 1.69 1.71 1.72 1.74 1.77 1.79
3.78 3.86 3.84 3.82 3.79 3.77 3.75 3.75
4.01 4.00 4.00 4.00 4.00 4.02 4.03 4.06 4.08 4.13 4.16 4.21
6.86 6.98 6.95 6.92 6.87 6.85 6.84 6.83 6.84 6.85 6.86 6.88
7.00* 7.11 7.08 7.06 7.01 6.98 6.97 6.97
7.41 7.53 7.50 7.47 7.43 7.40 7.38 7.37
9.18 9.46 9.40 9.33 9.23 9.14 9.07 9.01 8.96 8.92 8.89 8.85
10.01 10.32 10.25 10.18 10.06 9.97 9.89 9.83
12.46 13.42 13.21 13.01 12.64 12.30 11.99 11.71
* Non-NIST Phosphate Buffer
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Effects of Temperature – Sample Effects
• Sample effects
• Temperature compensation corrects for
changes in electrode slope not sample pH
• It is not possible to normalize pH readings to a
specific temperature
• pH of samples will change with temperature
changes
• Record temperature with pH readings
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Common Questions – Stable Readings
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Electrode Care and Maintenance
• Electrode Storage
• Short-term storage
• Use appropriate electrode storage solution. (ROSS or Standard)
• Alternatively, soak in 100 ml pH 7 buffer with 0.5 g KCl.
• Long-term storage
• Fill electrode, close fill hole, store with storage solution in protective cap
• Cleaning Solutions
• Soak electrode in solvent that will remove deposits
• Example: 0.1 M HCl for general cleaning
• Example: 1% pepsin in HCl for proteins
• Example: Bleach for disinfecting
• Example: detergent for grease & oil
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Electrode Care and Maintenance
• When do you need to clean your electrode?
• Check slope range
• Ideal range: 95% - 102%
• Cleaning range: 92% - 95%
• Replacement range: below 92%
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Electrode Care and Maintenance
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Common Questions - Maintenance
• Use cleaning remedies and cleaning solutions if you suspect a coated bulb or
coated junction is the cause of poor electrode slope.
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Keys to Accuracy
• Always use fresh buffers
• Check bottle expiration and date opened
• pH 4 and pH 7 buffers expire within
12 months of being opened.
• pH 10 buffer expires within 9 month of being
opened.
• Fresh buffer for each calibration
• Calibrate only once in buffer… don’t re-use
buffer
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Keys to Accuracy
• Make sure level of fill solution is
high
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Troubleshooting pH Problems
• Troubleshooting Sequence
• Meter
• Buffers
• Reference electrode
• pH electrode
• Sample
• Technique
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Troubleshooting pH Problems
• Troubleshooting pH Meters
• Use meter shorting strap
• Reading should be 0 mV +/- 0.2
mV
• Use meter self-test procedure
• Troubleshooting Buffers
• Use Fresh Buffers for calibration
• Verify expiration date
• 1 year after opening maximum
• Stir buffers during calibration
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Troubleshooting pH Problems
• Troubleshooting pH Electrodes
• Clean bulb, junctions
• Replace Fill solution
• Uncover fill hole
• Check for scratches on sensing bulb
• Troubleshooting Samples
• Proper sample preparation
• Stir samples
• Troubleshooting Technique
• Treat samples and buffers the same
• Clean and blot electrode between
samples
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Electrode Check
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Thermo Scientific – Technical Service
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Thank You!